Apparatus and methods for parallel processing of microvolume liquid reactions

Patent metadata

Field	Value
Publication number	US-10227644-B2
Application number	US-201815965155-A
Country	US
Kind code	B2
Filing date	Apr 27, 2018
Priority date	Feb 18, 2000
Publication date	Mar 12, 2019
Grant date	Mar 12, 2019

Abstract

Disclosed herein are apparatuses and methods for conducting multiple simultaneous micro-volume chemical and biochemical reactions in an array format. In one embodiment, the format comprises an array of microholes in a substrate. Besides serving as an ordered array of sample chambers allowing the performance of multiple parallel reactions, the arrays can be used for reagent storage and transfer, library display, reagent synthesis, assembly of multiple identical reactions, dilution and desalting. Use of the arrays facilitates optical analysis of reactions, and allows optical analysis to be conducted in real time. Included within the invention are kits comprising a microhole apparatus and a reaction component of the method(s) to be carried out in the apparatus.

First claim

What is claimed:

1. A system for conducting a plurality of polynucleotide reactions, the system comprising:
a substrate comprising an upper face, a lower face, and a plurality of holes disposed between the upper face and the lower face;
a dispenser that dispenses (1) liquid samples comprising nucleic acids, (2) reagents, and (3) oil into the plurality of holes;
a bath comprising an oil, the lower face of the substrate being submerged in the oil;
a thermal cycler that performs amplification reactions on the liquid samples dispensed within the plurality of holes to produce amplified products in the plurality of holes.

2. The system of claim 1 , wherein the plurality of holes comprise a plurality of through-holes in the substrate.

3. The system of claim 1 , further comprising an evaporation barrier that seals the plurality of holes.

4. The system of claim 1 , wherein the dispenser dispenses the samples in the plurality of holes via the upper face of the substrate.

5. The system of claim 1 , wherein the system creates sample libraries within the plurality of holes of the substrate.

6. The system of claim 1 , wherein the lower face of the substrate is immersed in the oil and the oil coats the samples.

7. The system of claim 1 , wherein the the lower face of the substrate is immersed in the oil to submerge, overlay, or coat the samples with the oil.

8. The system of claim 1 , wherein the thermal cycler comprises the bath.

9. The system of claim 1 , wherein the thermal cycler thermally cycles at least one of the oil or the bath.

10. The system of claim 1 , wherein the oil comprises mineral oil or silicone oil.

11. The system of claim 1 , wherein the substrate comprises a plate.

12. The system of claim 1 , wherein the thermal cycler comprises a heat block in thermal contact with the substrate and thermally cycles the liquid samples with the heat block.

13. The system of claim 1 , further comprising a sequencer, wherein the system produces an amplified product for loading into the sequencer.

14. The system of claim 1 , wherein the dispenser is a pipette.

15. A system for conducting a plurality of polynucleotide reactions, the system comprising:
a substrate comprising an upper face, a lower face, and a plurality of holes disposed between the upper face and the lower face;
a dispenser that dispenses (1) liquid samples comprising nucleic acids and (2) reagents into the plurality of holes, and a pipette that dispenses oil into the plurality of holes, the dispenser including an apparatus comprising samples arranged in a pattern matching a pattern of the plurality of holes;
a bath comprising an oil, the lower face of the substrate being submerged in the oil; and
a thermal cycler that performs amplification reactions on the liquid samples dispensed within the plurality of holes to produce amplified products in the plurality of holes.

16. The system of claim 15 , wherein the lower face of the substrate is immersed in the oil to submerge, overlay, or coat the samples with the oil.

17. The system of claim 15 , wherein the oil comprises mineral oil or silicone oil.

18. The system of claim 15 , wherein the substrate comprises a plate.

19. The system of claim 15 , further comprising a sequencer, wherein the system produces an amplified product for loading into the sequencer.

20. The system of claim 15 , wherein first dispenser is a first pipette and the second dispenser is a second pipette.

Assignees

• Univ Leland Stanford Junior

Inventors

• O'Keefe Matthew T
• Foreman Pamela K

Classifications

• B01J2219/00722

  Nucleotides · CPC title

• C12Q2600/172

  Haplotypes · CPC title

• B01F13/0071

  Operations & Transport · mapped topic

• B01L2300/087

  Multiple sequential chambers · CPC title

• C12Q1/6883

  for diseases caused by alterations of genetic material · CPC title